Abstract Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution high performance liquid chromatographic (HPLC) separations of larger molecules including proteins, peptides, glycopeptides, and glycans. Proteins therapeutics are complex molecules, susceptible to intended or unintended alterations of composition and conformation. Many therapeutic proteins are inherently a mixture of closely related variants. Current practice is to employ HPLC to resolve and quantify the components such protein mixtures, to assure identity and safety of the protein drug. Many HPLC separations of proteins are limited in resolution of protein mixtures, require an hour or more, and may not completely resolve all of the components. Due to the complexity of samples, there are many examples in the current research literature that combine multiple HPLC analyses to gain full details on protein composition, and to measure the levels of protein variants, both during development of the drug, as well as manufacture and quality assurance/quality control. Regardless of the intended use of the quantitative information, high efficiency LC separations are a fundamental part of the analytical systems, and the time to achieve high resolution separations of protein samples is a great bottleneck. This proposal describes an approach to improve the separation efficiency of HPLC for protein analysis by extending the use of novel superficially porous particle (SPP) technologies to larger pore size column packing materials of smaller size particles, with optimized characteristics, providing significantly faster and higher resolution separations of proteins. Our objectives are to create 2.0 ?m diameter and smaller SPP silica particles, optimize their materials properties, and to load these materials efficiently into HPLC column formats. The proposed work will extended recent breakthroughs in material science that yield authentic improvements in current materials, seeking to employ the technology in additional modes of HPLC operation, including hydrophobic interaction, ion exchange, and hydrophilic interaction. The goals of the proposed work would yield very high performance chromatographic products, greater than those currently available, to be applied broadly in analysis of proteins, particularly for protein therapeutics. The application of the technology is not limited to biopharmaceutical preparations, and, in fact, would benefit any current application that uses HPLC methods for larger biological molecule analyses. The separations technology described will directly lead to useful products for which there is a significant technical and market demand.